Salt-tolerant bacillus amyloliquefaciens with high self-aggregation capability and application thereof in denitrification

A technology of amyloliquefaciens and bacillus, applied in the field of environmental microorganisms, can solve the problems of low denitrification efficiency and achieve high denitrification performance, high self-aggregation ability, and high removal rate

Active Publication Date: 2021-08-13
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The traditional biological nitrogen removal method needs to separate the aerobic and anoxic phases, resulting in low nitrogen removal efficiency

Method used

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  • Salt-tolerant bacillus amyloliquefaciens with high self-aggregation capability and application thereof in denitrification
  • Salt-tolerant bacillus amyloliquefaciens with high self-aggregation capability and application thereof in denitrification
  • Salt-tolerant bacillus amyloliquefaciens with high self-aggregation capability and application thereof in denitrification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The screening of bacterial strain N8 is carried out according to the following steps:

[0030] (1) Take high-density grouper culture water in a 50ml sterile centrifuge tube and store at 4°C for later use. Take 1mL water sample and put it into a 250mL Erlenmeyer flask, add 100mL nitrification enrichment medium; shake well, 30℃, 180r min －1 After 2 days, take 1 mL and re-inoculate it into fresh nitrification-enrichment medium, and then enrich and culture continuously for 5 times.

[0031] (2) Under aseptic conditions, use a pipette to draw 1 mL of the above-mentioned last-generation liquid-enriched culture solution, add it to a 20 mL test tube filled with 9 mL of sterile water, rinse with a pipette for 3 times, and mix well , which makes 10 －1 Concentration of bacterial liquid, repeat the above operations, and finally made 10 －3 、10 －4 、10 －5 、10 －6 and 10 －7 Bacterial solutions with different dilutions. Use a pipette gun to draw 0.1mL of bacterial solutions with ...

Embodiment 2

[0040] Characteristic and Identification of Strain N8

[0041] The colony of strain N8 is smaller, moister, smoother, more transparent, easy to pick, with neat edges, smooth and shiny surface. Under the microscope, the bacteria are rod-shaped and are Gram-negative bacteria. Its relevant physiological and biochemical characteristics are shown in Table 1, V-P test, gelatin liquefaction, starch hydrolysis test are positive; nitrate reduction test is negative, citrate, propionate, L-arabinose, D-mannitol can be used as Carbon source, D-xylose cannot be utilized. It cannot grow in 7% NaCl, but can grow under pH 5.7 and anaerobic conditions.

[0042] Table 1 Physiological and biochemical characteristics of strain N8

[0043]

[0044] +: growing or positive; -: not growing or negative

[0045] 16S rDNA gene sequencing and multiplex PCR identification:

[0046] PCR amplification of strain N8 genomic DNA: the total DNA of strain N8 was used as a template, and the amplification ...

Embodiment 3

[0056] Salt Tolerance Test of Bacillus amyloliquefaciens N8

[0057] Use an inoculation loop to scrape 2 loops of bacterial lawn from Bacillus amyloliquefaciens N8 stored on the inclined tube medium, inoculate it into a 250mL Erlenmeyer flask containing 100mL of sterilized LB medium, at 30°C, 140r min －1 Cultivated under the condition, until the medium OD 600 When it is 1, it is used as seed solution.

[0058] Prepare nitrification basal medium with different salinity gradients, in which NaCl concentrations are 0, 10‰, 20‰, 30‰, 40‰ and 50‰ (W / V); inoculate Bacillus amyloliquefaciens N8 seed solution with 1% (v / v) Transfer to a 500mL Erlenmeyer flask containing 300mL nitrification basal medium; shake well, 30°C, 140r min －1 Cultivate for 48 hours under the same conditions, and the strain growth status (OD 600 ) as the basis of whether the strain is resistant to the corresponding salinity; the culture medium not inoculated with bacteria is used as the blank control, and eac...

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Abstract

The invention discloses a salt-tolerant bacillus amyloliquefaciens with high self-aggregation capability and application thereof the in denitrification. The bacillus amyloliquefaciens N8 has been preserved in the Guangdong Microbial Culture Collection Center on June 1, 2021, and the preservation number of the bacillus amyloliquefaciens N8 is GDMCC NO.61606. The bacillus amyloliquefaciens N8 can be applied to denitrification of nitrogen-containing wastewater with certain salinity. The bacillus amyloliquefaciens N8 has the nitrification capability of efficiently removing ammonia nitrogen and the denitrification capability of removing nitrite and nitrate nitrogen; the bacillus amyloliquefaciens N8 has certain salt tolerance, and can effectively exert the denitrification capability in a salt-containing condition. By utilizing the bacteria with salt-tolerant nitrification and denitrification capabilities, the problem of removing ammonia nitrogen, nitrite and nitrate nitrogen in wastewater under a salt-containing condition can be well solved.

Description

technical field [0001] The invention belongs to the field of environmental microorganisms, and in particular relates to a strain of bacillus amyloliquefaciens with high self-aggregation ability and salt tolerance and its application in denitrification. Background technique [0002] At present, with the development of the aquaculture industry to a modern level, the recirculating aquaculture system (RAS) has gradually become a supporting technical force for the modernization of China's aquaculture industry due to its small footprint, water conservation, and environmental protection. Good water quality is the technical core and key advantage of RAS, and the concentration of ammonia nitrogen is the most critical factor affecting water quality. In addition, the harm caused by the accumulation of nitrite nitrogen in the breeding process is also serious. At the same time, fatal or near-fatal effects of high concentrations of nitrate on marine and freshwater organisms have also bee...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/07C02F101/16
CPCC02F3/34C02F2101/16C02F2101/163C02F2101/166C12N1/20
Inventor 叶超霞刘博张文倩
Owner SOUTH CHINA NORMAL UNIVERSITY
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